Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
Moscow State University, Moscow, Russia.
Dokl Biochem Biophys. 2022 Apr;503(1):90-92. doi: 10.1134/S1607672922020077. Epub 2022 May 10.
Seven amino acid sequences of antibody mimetics molecules, monobodies, capable of interacting with the nucleocapsid protein of the SARS-CoV virus, were taken from the literature. Nucleotide sequences of monobody genes were obtained by gene synthesis, which were expressed in E. coli and isolated using Ni-NTA chromatography. It was shown by thermophoresis that three of the seven selected antibody-like molecules can interact with high affinity (dissociation constant of tens of nM) with the nucleocapsid protein of the SARS-CoV-2 virus. For the remaining four monobodies, only low affinity binding with a dissociation constant of several μM was found.
从文献中选取了 7 种能够与 SARS-CoV 病毒核衣壳蛋白相互作用的抗体模拟分子（单域抗体）的 7 个氨基酸序列。通过基因合成获得了单域抗体基因的核苷酸序列，在大肠杆菌中表达并通过 Ni-NTA 层析法进行分离。通过热迁移实验表明，在这 7 种选择的抗体样分子中，有 3 种能够与 SARS-CoV-2 病毒的核衣壳蛋白以高亲和力（解离常数为几十 nM）相互作用。对于其余的 4 种单域抗体，仅发现与解离常数为几个 μM 的低亲和力结合。
